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CNPY2 / Protein canopy homolog 2 · IHC design guide
Design Immunohistochemistry for CNPY2
Plan CNPY2 chromogenic IHC around the widespread cytoplasmic tissue pattern (HPA tissue IHC) and its ER localization (UniProt). Use the catalog antibody’s 1:50–1:200 IHC dilution range as a starting point (datasheet A10020), and interpret staining with the reported multi-gene targeting caution (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
CNPY2 Immunohistochemistry Experimental Design Guide
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before staining
| Expected localisation | Cytoplasmic staining (HPA tissue IHC) | |
| Staining pattern | Widespread cytoplasmic staining across cell types (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, heat-mediated (datasheet A10020) | |
| Positive control | Cervix+4 more · see all | |
| Negative control | Oral mucosa |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep paraffin-section fixation consistent (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A10020) | |
| Caveat | Staining may reflect protein from more than one gene (HPA tissue IHC) | |
| Regulation | Staining intensity varies by tissue (HPA tissue IHC) | |
| Isoform / epitope | 2 isoforms; signal peptide 1–20 is cleaved (UniProt) |
Section 1
Recommended CNPY2 IHC & IF Protocols
The catalog antibody protocol is followed by three published chromogenic IHC protocols for CNPY2 in liver, myocardium, and tissue sections (PMC8306219; PMC6443342; PMC4230931).
| Sample | Paraffin-embedded mouse kidney tissue; fixative not specified (datasheet A10020) |
| Fixation | Image fixative and duration unreported (datasheet A10020); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Citrate pH 6.0 (datasheet A10020); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit anti-CNPY2, 1:50-1:200 (datasheet A10020) |
| Primary incubation | Overnight at 4 °C (standard) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | CNPY2-positive staining in glandular cells of cervix (HPA tissue IHC: High). HPA tissue profile: Ubiquitous cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteStart with heat retrieval in citrate at pH 6.0, as specified for this page and used in two published protocols (datasheet A10020; PMC8306219; PMC4230931).
Section 2
What Is the Expected CNPY2 Staining Pattern?
CNPY2 should appear predominantly cytoplasmic in tissue IHC (HPA: ubiquitous cytoplasmic expression), consistent with an endoplasmic reticulum annotation and no transmembrane segment (UniProt Q9Y2B0: location and topology). Expect strong staining in placental trophoblastic cells and in glandular cells of the cervix, epididymis, fallopian tube and thyroid (HPA: High). HPA rates its tissue pattern Supported, while cautioning that the staining may represent proteins from more than one gene (HPA: reliability).
| Cytoplasmic staining in placental trophoblastic cells or the listed glandular cells. | This matches the reported High tissue IHC pattern (HPA: tissue IHC). Compare cell types and compartments before calling a section positive; intensity alone cannot resolve HPA's multi-gene staining caveat (HPA: reliability). |
| Predominantly nuclear staining, with little cytoplasmic signal. | That compartment does not match the reported cytoplasmic tissue pattern or the endoplasmic reticulum annotation (HPA: tissue IHC; UniProt Q9Y2B0: location). Check staining and counterstain interpretation before assigning it to CNPY2 (general IHC practice). |
| Strong signal in oral mucosal squamous epithelium. | HPA reports CNPY2 as Not detected in these cells (HPA: oral mucosa). Unexpected staining warrants a specificity check; cross-reactivity or endogenous chromogen activity are possibilities, not diagnoses from appearance alone (general IHC practice). |
| Diffuse colour across cells and surrounding tissue, without a clear cellular pattern. | The pattern is hard to reconcile with HPA's cytoplasmic cell staining (HPA: tissue IHC). Review background in a no-primary control and inspect blocking, washing and detection steps (general IHC practice). |
| No staining in placental trophoblastic cells despite an apparently intact section. | This conflicts with HPA's High report for that cell type (HPA: placenta). Check section identity, antibody and detection performance, and controls before interpreting other unstained tissues as negative (general IHC practice). |
💡Expected CNPY2 appearanceCall a result positive when cytoplasmic signal is strong in placental trophoblastic cells or the listed glandular cells (HPA: High; cytoplasmic profile); predominant nuclear colour or indiscriminate tissue-wide colour calls for review (general IHC practice).
| Tissue and cell choice | Placental trophoblastic cells and the listed glandular cells offer reported High staining; oral mucosal squamous cells are reported Not detected (HPA: tissue IHC). These are cell-specific comparisons, not guarantees for every section (general IHC practice). |
| Antibody evidence | HPA038465 and HPA038466 each have Supported IHC status (HPA: antibodies). HPA also cautions that tissue staining may target proteins from more than one gene, so a matching pattern alone does not establish unique CNPY2 detection (HPA: reliability). |
| Protein location and processing | UniProt places CNPY2 in the endoplasmic reticulum, finds no transmembrane segment, and records a signal peptide at residues 1–20 followed by a 21–182 chain (UniProt Q9Y2B0). The antibody epitope is unspecified; these annotations cannot predict its staining performance. |
| Isoforms | UniProt records 2 CNPY2 isoforms (UniProt Q9Y2B0: isoforms). The payload does not map an antibody epitope to either isoform, so staining intensity cannot be assigned to a particular isoform. |
| IF/ICC Q&A: should its pattern match tissue IHC? | HPA reports approved cytosolic ICC-IF localisation and images from A-431, U-251MG and U2OS (HPA: subcellular; HPA038465: ICC Approved). Tissue IHC reports a cytoplasmic profile (HPA: tissue IHC); compare compartment, while evaluating IF/ICC on its own guide page. |
| Situation | Likely cause | Next action |
|---|---|---|
| Placental trophoblastic cells show no visible chromogenic signal (HPA: placenta, High). | A failed staining or detection step is possible; the image alone cannot identify which step failed (general IHC practice). | Confirm the cell type, inspect a positive control, then review antibody application, retrieval and detection records (general IHC practice). |
| Most colour is nuclear, with little cytoplasmic staining. | This conflicts with the reported cytoplasmic IHC profile and endoplasmic reticulum annotation (HPA: tissue IHC; UniProt Q9Y2B0: location). | Review counterstain and chromogen channels, then repeat with appropriate staining controls if the compartment remains unclear (general IHC practice). |
| Oral mucosal squamous epithelium stains strongly (HPA: Not detected). | Unexpected signal could reflect nonspecific antibody binding or endogenous detection activity; HPA's multi-gene caution limits attribution (HPA: reliability; general IHC practice). | Compare a no-primary control and the expected positive cell types in the same run; reassess specificity before scoring (general IHC practice). |
| Colour is diffuse across cells and tissue spaces. | Background from detection or washing is possible; diffuse colour does not show HPA's cell-associated cytoplasmic profile (HPA: tissue IHC; general IHC practice). | Check the no-primary control, blocking and wash steps, then reassess whether cellular boundaries and cytoplasm can be scored (general IHC practice). |
| Positive cells stain, but their intensity varies among sections. | Different cell populations may have different reported levels; HPA records High, Medium and Low categories across specified tissues and cells (HPA: tissue IHC). | Compare like cell types under the same scoring conditions and check run controls before interpreting a weaker section (general IHC practice). |
| An apparently plausible cytoplasmic pattern is being called CNPY2-specific. | HPA calls its IHC evidence Supported but cautions that staining may represent proteins from more than one gene (HPA: reliability). | Report the observed cell type, compartment and intensity separately from target attribution; use an independent specificity check if the conclusion requires unique CNPY2 detection (general IHC practice). |
Experiment evidence
IHC & IF images showing CNPY2 staining
A10020 · IHCImmunohistochemistry of paraffin-embedded mouse kidney using CNPY2 Rabbit pAb at dilution of 1:100 .Perform high pressure antigen retrieval
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000257727-CNPY2/tissue/cerebral+cortex#imid_9985537
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000257727-CNPY2/tissue/colon#imid_9985455
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000257727-CNPY2/tissue/liver#imid_9985464
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000257727-CNPY2/tissue/kidney#imid_9985465
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA038465; green = CNPY2, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA038465; green = CNPY2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for CNPY2 IHC & IF
🧪Run placenta first and expect staining in trophoblastic cells (HPA: placenta, trophoblastic cells High). Run oral mucosa as the negative tissue (HPA: oral mucosa, squamous epithelial cells Not detected); within the placenta slide, compare trophoblastic staining with morphologically distinct areas at background level, without assuming those cells are CNPY2-negative.
Positive control tissue: Cervix (Glandular cells, HPA High)
Negative control tissue: Oral mucosa (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show CNPY2 in A-431, U-251MG, U2OS, with annotated localisation: Cytosol (approved) (HPA subcellular).
Technical controls: Run a no-primary (secondary-only) control, a concentration-matched nonimmune rabbit IgG control appropriate to the catalog rabbit polyclonal antibody, and CNPY2 knockout material if available or a cognate peptide-block control if the immunogen is known (A10020 tissue-IHC caption: Rabbit pAb; standard IHC practice). For placental chromogenic sections, quench endogenous peroxidase and check blood-associated background; if using IF, inspect tissue autofluorescence (standard IHC/IF practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A10020 paraffin-section caption does not state a fixative (A10020 tissue-IHC caption). That caption uses high-pressure retrieval in 10 mM citrate buffer at pH 6.0 and a 1:100 antibody dilution, but does not establish whether retrieval is required (A10020 tissue-IHC caption). The supplied evidence does not establish that frozen sections or IF are easier; interpret IF localisation cautiously because HPA reports cytosol in ICC-IF whereas UniProt assigns CNPY2 to the endoplasmic reticulum (HPA: subcellular; UniProt Q9Y2B0: subcellular location).
HPA tissue IHC evidence for CNPY2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — High consistency between antibody staining and RNA expression data. Caution, targets protein from more than one gene.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Cervix | Glandular cells | High | Protein (IHC) | HPA → |
| Epididymis | Glandular cells | High | Protein (IHC) | HPA → |
| Fallopian tube | Glandular cells | High | Protein (IHC) | HPA → |
| Placenta | Trophoblastic cells | High | Protein (IHC) | HPA → |
| Thyroid gland | Glandular cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Oral mucosa | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced CNPY2 IHC Tips
Troubleshoot CNPY2 staining in paraffin sections by checking retrieval, compartment, cell identity, and controls before comparing staining intensity.
What retrieval conditions should I start with for CNPY2 in paraffin sections?
Start with heat-mediated retrieval in 10 mM citrate, pH 6.0, for paraffin sections (datasheet A10020; A10020 kidney caption). The mouse kidney image used high-pressure retrieval and an antibody dilution of 1:100; its fixative was unreported (A10020 kidney caption). Keep heating and cooling conditions identical across slides, then compare staining with a no-primary control and an expected-positive section (standard IHC practice; HPA tissue IHC). If staining remains weak, optimize heating duration before testing another retrieval buffer, because excessive heat can damage morphology (standard IHC practice). Record retrieval conditions alongside staining intensity so a stronger signal is not mistaken for greater CNPY2 abundance (standard IHC practice).
How should I troubleshoot fixation-related loss of CNPY2 staining?
Target-specific fixation sensitivity is unknown: the selected paraffin-section caption does not report a fixative, and it cannot establish an FFPE result (A10020 kidney caption). When staining varies between blocks, document fixative, fixation duration, tissue thickness, and processing history before changing antibody conditions (standard IHC practice). Compare sections processed together using the same 10 mM citrate, pH 6.0 retrieval and the caption’s 1:100 starting dilution (datasheet A10020; A10020 kidney caption). Include an expected-positive control on each run, then assess whether signal loss tracks processing batches or tissue regions (HPA tissue IHC; standard IHC practice). Do not assign a CNPY2-specific fixation effect without a direct comparison (A10020 kidney caption).
Where should CNPY2 staining appear, and how should conflicting patterns be assessed?
Assess intracellular staining across the cell body: UniProt assigns CNPY2 to the endoplasmic reticulum, whereas HPA reports approved cytosolic localisation and ubiquitous cytoplasmic tissue staining (UniProt Q9Y2B0; HPA subcellular; HPA tissue IHC). Compare a granular or diffuse cytoplasmic pattern with tissue architecture and a no-primary control before assigning an organelle (standard IHC practice). A predominantly nuclear, cell-surface, or extracellular chromogenic pattern warrants scrutiny against these reported locations (UniProt Q9Y2B0; HPA subcellular). Because CNPY2 has a cleavable 1–20 signal sequence and no transmembrane segment, topology alone cannot identify the antibody’s stained compartment (UniProt Q9Y2B0). Document the actual pattern rather than forcing agreement between the two localisation resources (UniProt Q9Y2B0; HPA subcellular).
Could isoforms or epitope accessibility explain different CNPY2 staining patterns?
CNPY2 has 2 listed isoforms, but the supplied product caption does not identify the antibody epitope or establish isoform recognition (UniProt Q9Y2B0; A10020 kidney caption). Record which antigen region a separate antibody recognizes before comparing section staining or claiming isoform specificity (standard IHC practice). The precursor has a 1–20 signal peptide, a 21–182 mature chain, a Saposin B-type domain, and phosphoserine 115 (UniProt Q9Y2B0). Those features suggest regions to examine when epitope information becomes available; they do not establish which feature affects retrieval (UniProt Q9Y2B0). Compare antibodies against documented distinct epitopes on matched sections if an isoform-dependent interpretation is essential (standard IHC practice).
How can IF help check the cell type and compartment seen by chromogenic IHC?
Use IF as a separate validation experiment, pairing CNPY2 with a marker independently validated for the expected cell type, such as trophoblastic cells in placenta (HPA tissue IHC; standard IF practice). Choose spectrally separated fluorophores and check an unstained section for tissue autofluorescence before interpreting overlap (standard IF practice). CNPY2 is reported in the endoplasmic reticulum and cytosol, with a signal peptide and no transmembrane segment (UniProt Q9Y2B0; HPA subcellular). Permeabilise for intracellular access, then confirm the antibody epitope’s membrane-facing side from epitope data rather than assuming it from topology (standard IF practice; UniProt Q9Y2B0). Include single-label and no-primary controls when assessing apparent colocalisation (standard IF practice).
What should I check when CNPY2 chromogenic staining is widespread or muddy?
Widespread cytoplasmic signal can be plausible because HPA describes ubiquitous expression, but its tissue-IHC assessment also cautions that the staining may detect protein from more than one gene (HPA tissue IHC). Check no-primary and reagent controls, then inspect whether signal persists where specific antibody binding should be absent (standard IHC practice). Quench endogenous peroxidase for an enzyme-based chromogenic workflow and compare with a control processed through the same DAB steps (standard IHC practice). If background remains high, optimize antibody incubation and blocking while holding retrieval at 10 mM citrate, pH 6.0 initially (datasheet A10020; standard IHC practice). Judge changes against preserved cellular detail and an expected-positive section, not darkness alone (HPA tissue IHC; standard IHC practice).
How should I quantify CNPY2 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the scored cell population and anatomical region before measuring CNPY2, because HPA reports different staining levels across cell types and tissues (HPA tissue IHC). For cell-level chromogenic signal, report an H-score from intensity and percentage of positive cells; for a defined population, report percentage positive (standard IHC practice). If counting positive cells, express density per mm² of viable tissue or the prespecified compartment, and exclude folds and necrosis (standard IHC practice). Keep retrieval, antibody exposure, detection time, and image settings consistent across compared slides (standard IHC practice). Normalise to the same eligible cell population or tissue area, and include an expected-positive control for each staining run (HPA tissue IHC; standard IHC practice).
How can I distinguish convincing CNPY2 staining from tissue artefact?
A convincing result places signal in identifiable cell cytoplasm, consistent with reported endoplasmic-reticulum and cytosolic locations, while retaining the expected tissue architecture (UniProt Q9Y2B0; HPA subcellular; standard IHC practice). Compare the stained cell type with HPA examples, including high trophoblastic-cell staining in placenta and undetected squamous epithelial staining in oral mucosa (HPA tissue IHC). Flag staining confined to cut edges, folds, or necrotic areas as suspect and compare it with adjacent intact tissue (standard IHC practice). Evaluate nuclear or exclusively extracellular signal against the reported locations, and rule out endogenous enzyme colour with a no-primary control (UniProt Q9Y2B0; HPA subcellular; standard IHC practice). Treat agreement with HPA cautiously because its tissue-IHC assessment warns of detection from more than one gene (HPA tissue IHC).
Boster reagents
Best CNPY2 / Protein canopy homolog 2 IHC Antibodies
A10020 has a real IHC image of paraffin-embedded mouse kidney (IHC caption). Its listed reactivity covers human, mouse and rat; no IF/ICC data are provided (catalog: A10020).
Real IHC data
Anti-Protein canopy homolog 2 CNPY2 Antibody
Cat # A10020
A10020 is shown staining paraffin-embedded mouse kidney at 1:100 after high-pressure retrieval in 10 mM citrate, pH 6.0 (IHC caption). Its listed applications include IHC, and its listed reactivity covers human, mouse and rat; the payload provides no IF image (catalog: A10020).
Which to pick: Choose A10020 for paraffin-section IHC because its own image documents staining in mouse kidney, while the fixative is unreported (IHC caption). No listed SKU has IF/ICC validation, so there is no supported IF/ICC pick here (catalog: A10020). A10020 is a rabbit polyclonal antibody with listed human, mouse and rat reactivity; its pictured IHC evidence is limited to mouse kidney (catalog: A10020; IHC caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9Y2B0 (CNPY2_HUMAN, Protein canopy homolog 2).
- Human Protein Atlas. CNPY2 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. CNPY2 subcellular location (ICC-IF): Localized to the cytosol..
- Human Protein Atlas. CNPY2 antibody validation summary (2 antibodies).
- Canopy Homolog 2 as a Novel Molecular Target in Hepatocarcinogenesis. Cancers 2021 — PMC8306219.
- Protective effect of Barbaloin in a rat model of myocardial ischemia reperfusion injury through the regulation of the CNPY2‑PERK pathway. International journal of molecular medicine 2019 — PMC6443342.
- Expression of CNPY2 in mouse tissues: quantification and localization. PloS one 2014 — PMC4230931.
- CNPY2 inhibits MYLIP-mediated AR protein degradation in prostate cancer cells. Oncotarget 2018 — PMC5915145.
- PubMed PMID:10072769 — UniProt-cited evidence.
- PubMed PMID:12826659 — UniProt-cited evidence.
- PubMed PMID:12975309 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for CNPY2 (Protein canopy homolog 2). Expected localisation Cytoplasmic staining (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, heat-mediated (datasheet A10020). Validated IHC antibody: A10020 at 1:50-1:200 (the product's own IHC validation image is shown). Controls (HPA): positive Cervix (Glandular cells); negative Oral mucosa. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded mouse kidney tissue; fixative not specified (datasheet A10020); 2. Fixation: Image fixative and duration unreported (datasheet A10020); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat retrieval: Citrate pH 6.0 (datasheet A10020); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit anti-CNPY2, 1:50-1:200 (datasheet A10020); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep paraffin-section fixation consistent (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A10020); Caveat: Staining may reflect protein from more than one gene (HPA tissue IHC); Isoform / epitope: 2 isoforms; signal peptide 1–20 is cleaved (UniProt); Regulation: Staining intensity varies by tissue (HPA tissue IHC). Evidence: UniProt Q9Y2B0, HPA tissue IHC · Supported, HPA ICC-IF · cytosol., Datasheet A10020, PMC8306219, PMC6443342. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/CNPY2/immunohistochemistry-ihc-guide.